Genetics and Molecular Biology, Volume: 28, Issue: 1, Published: 2005
  • Phage display technology: applications and innovations Human And Medical Genetics

    Arap, Marco Antonio

    Abstract in English:

    The expression of exogenous peptides on the surface of filamentous bacteriophage was initially described by Smith in 1985. Since his first study, different molecules such as small peptides and antibodies have been displayed on coat proteins of phage, greatly expanding the applications of the technology. The past decade has seen considerable progress in the techniques and applications of phage libraries. In addition, different screening methods have allowed isolation and characterization of peptides binding to several molecules in vitro, in the context of living cells, in animals and in humans. Here we review the applications, recent innovations, and future directions of phage display technology.
  • Structure and stability upon maternal transmission of common and intermediate FMR1 (Fragile X Mental Retardation 1) alleles in a sample of the Brazilian population Human And Medical Genetics

    Capelli, Leonardo P.; Mingroni-Netto, Regina C.; Vianna-Morgante, Angela M.

    Abstract in English:

    In order to investigate the stability of the FMR1 (Fragile X Mental Retardation 1) alleles from the normal population, when maternally inherited, we analyzed 75 mother-to-son transmissions. Sixty-eight alleles fell within the common range with 20-40 CGG repeats, and seven alleles were intermediate, with 41-48 repeats. No change was observed either in the length or in the structure of these repeats upon transmission. Fifty-three alleles were ascertained in different families, and their size distribution was similar to those described for European and European-derived populations, with three peaks of frequency: 66% of the alleles with (CGG)29, (CGG)30 or (CGG)31, 7.5% with (CGG)20, and 5.7% with (CGG)23. Regarding the AGG interspersion pattern, 69.8% had two AGG repeats, 20.8% had one, 5.7% had three and 3.8% had none. The most common patterns were 10+9+9 (30.2%), 9+9+9 (18.9%), 10+9 (7.5%), and 10+9+10 (7.5%). About 70% of the alleles with up to 40 repeats were linked to the DXS548/FRAXAC1 haplotype 7-3, the most commonly reported in normal populations. Four out of five intermediate alleles were in linkage with the two haplotypes most frequently associated to the FMR1 full mutation, 2-1 and 6-4. These four alleles showed long uninterrupted CGG repeats at the 3' end. The 9+9+22, 9+9+23 and 9+9+28 alleles were linked to the haplotype 2-1, and the 9+37 allele, to the haplotype 6-4. The pattern of AGG interspersion of these alleles and the associated haplotypes were in accordance with the two main pathways toward mutation previously proposed.
  • Association of a PIT1 gene polymorphism with growth hormone mRNA levels in pig pituitary glands Human And Medical Genetics

    Franco, Maurício Machaim; Antunes, Robson Carlos; Oliveira, Katiana Mello de; Pereira, Cícero Donizete; Biase, Fernando Henrique; Nunes, Francis de Morais Franco; Goulart, Luiz Ricardo

    Abstract in English:

    Fourty-six non-castrated, halothane-free, male Landrace pigs were genotyped by PCR-RFLP for the Rsa I polymorphism in the PIT1 gene and classified into AA and AB genotypes. Total RNA was extracted from the pituitaries and the relative quantities of growth hormone (GH) mRNA were determined by semi-quantitative RT-PCR. Pigs with the AB genotype had higher levels of GH mRNA than those with the AA genotype (p = 0.034; Kruskal-Wallis test). This result suggests that the Rsa I polymorphism may be involved in Pit-1 protein expression or function, which in turn may influence GH transcription and expression. Thus, the Rsa I PIT1 gene polymorphism in this pig line may be used as a molecular marker to identify higher GH expression and possibly select for carcass and performance traits affected by GH.
  • Genetic characterization of the population of São Luís, MA, Brazil Human And Medical Genetics

    Ferreira, Francileide Lisboa; Leal-Mesquita, Emygdia Rosa; Santos, Sidney Emanuel Batista dos; Ribeiro-dos-Santos, Ândrea Kely Campos

    Abstract in English:

    Five loci (vWA1, F13A1, D12S67, Apo-B and D1S80) were investigated by polyacrylamide gel electrophoresis followed by silver staining in a sample of 177 individuals from the population of São Luís, State of Maranhão, Brazil. A total of 70 different alleles were identified. A statistically significant deviation from the Hardy-Weinberg equilibrium was observed in a single locus (F13A1, p = 0.0075). The average heterozygosity (H) was estimated at 77.7%, the mean number of alleles per locus as 14. The PD (capacity of genotype differentiation at each locus) ranged from 88.9% (vWA1) to 96.7% (F13A1). The combined PE (power of exclusion) of these five loci was 99.8%. In terms of racial admixture (42% European, 39% Indian, and 19% African Black ancestry), São Luís presented an estimate similar to Belém, another trihybrid Amazonian population.
  • Distribution of glutathione S-transferase GSTM1 and GSTT1 null phenotypes in Brazilian Amerindians Human And Medical Genetics

    Klautau-Guimarães, Maria de Nazaré; Hiragi, Cássia de Oliveira; D'Ascenção, Renata Ferreira; Oliveira, Silviene Fabiana; Grisolia, Cesar Koppe; Hatagima, Ana; Ferrari, Íris

    Abstract in English:

    The distribution of glutathione S-transferase (GST) GSTM1 and GSTT1 null phenotype frequencies in two Brazilian Amerindian tribes, the Munduruku tribe from Missão Cururu village (79 individuals) and the Kayabi tribe (41 individuals), was analyzed by polymerase chain reaction (PCR) amplification. The GST null phenotype frequencies for the Munduruku sample were 0% for GSTM1 and 27% for GSTT1 while for the Kayabi sample the null phenotype frequencies were 27% for GSTM1 and 29% for GSTT1. This is the first report of the absence of the GSTM1 null phenotype in any ethnic group.
  • Frequency of the Q192R and L55M polymorphisms of the human serum paraoxonase gene (PON1) in ten Amazonian Amerindian tribes Human And Medical Genetics

    Santos, Ney Pereira Carneiro dos; Ribeiro-dos-Santos, Ândrea KC; Santos, Sidney EB

    Abstract in English:

    Human serum paraoxonase (PON1) is an esterase associated with high density lipoproteins (HDLs) in the plasma and may confer protection against coronary artery disease. Serum PON1 levels and activity vary widely among individuals and populations of different ethnic groups, such variations appearing to be related to two coding region polymorphisms (L55M and Q192R). Several independent studies have indicated that the polymorphism at codon 192 (the R form) is a significant risk factor for cardiovascular disease in some populations, although this association has not been confirmed in other populations. Given the possible associations of these mutations with heart diseases and the fact that little or nothing is known of their prevalence in Amerindian populations, we investigated the variability of both polymorphisms in ten Amazonian Indian tribes and compared the variation found with that of other Asian populations in which both polymorphisms have been investigated. The results show that the LR haplotype is the most frequent and the MR haplotype is absent in all Amerindians and Asian populations. We also found that South America Amerindians present the highest frequency of the PON1192*R allele (considered a significant risk factor for heart diseases in some populations) of all the Amerindian and Asian populations so far studied.
  • Chromosome 17 abnormalities and mutation of the TP53 gene: correlation between cytogenetics, flow cytometry and molecular analysis in three cases of chronic myeloid leukemia Human And Medical Genetics

    Otero, Luize; Cavalcanti Júnior, Geraldo Barroso; Klumb, Claudete Esteves; Scheiner, Marcos Antonio Mauricio; Magluta, Eliane Pereira Simões; Fernandez, Teresa de Souza; Silva, Maria Luiza Macedo; Pires, Virgínia; Andrade, Gabriela Vasconcelos; Maia, Raquel Ciuvalschi; Tabak, Daniel

    Abstract in English:

    chronic myeloid leukemia (CML) have been described. This chromosomal region contains the tumor suppressor gene TP53 that may be an important factor in the evolution of this disease. In this study, we used flow cytometry and western blotting to assess p53 protein expression and single stranded conformational polymorphism to examine TP53 gene alterations in three patients with CML who showed alterations in 17p. Only the case with del(17)(p11) had p53 expression positive by flow cytometry and an abnormal migration pattern by SSCP analysis. The importance of the correlation between the results obtained with these techniques, as well as the clinical course of the patients, are discussed.
  • GATM, the human ortholog of the mouse imprinted Gatm gene, escapes genomic imprinting in placenta Human And Medical Genetics

    Miyamoto, Toshinobu; Sengoku, Kazuo; Hayashi, Hiroaki; Sasaki, Yoshihito; Jinno, Yoshihiro; Ishikawa, Mutsuo

    Abstract in English:

    The GATM gene encodes L-arginine:glycine amidinotransferase, which catalyzes the conversion of L-arginine into guanidinoacetate, the rate-limiting step in the synthesis of creatine. Since, deficiencies in creatine synthesis and transport lead to certain forms of mental retardation in human, the human GATM gene appears to be involved in brain development. Recently it has been demonstrated that the mouse Gatm is expressed during development and is imprinted with maternal expression in the placenta and yolk sac, but not in embryonic tissues. We investigated the imprinting status of the human GATM by analyzing its expression in four human placentas. GATM was biallelically expressed, thus suggesting that this gene escapes genomic imprinting in placentas, differently from what has been reported in mouse extra-embryonic tissues.
  • Molecular phylogeny of the genus Saguinus (Platyrrhini, Primates) based on the ND1 mitochondrial gene and implications for conservation Animal Genetics

    Tagliaro, Claudia Helena; Schneider, Horacio; Sampaio, Iracilda; Schneider, Maria Paula Cruz; Vallinoto, Marcelo; Stanhope, Michael

    Abstract in English:

    The systematics of the subfamily Callitrichinae (Platyrrhini, Primates), a group of small monkeys from South America and Panama, remains an area of considerable discussion despite many investigations, there being continuing controversy over subgeneric taxonomic classifications based on morphological characters. The purpose of our research was to help elucidate the phylogenetic relationships within the monkey genus Saguinus (Callitrichinae) using a molecular approach to discover whether or not the two different sections containing hairy-faced and bare-faced species are monophyletic, whether Saguinus midas midas and Saguinus bicolor are more closely related than are S. midas midas and Saguinus midas niger, and if Saguinus fuscicollis melanoleucus and Saguinus fuscicollis weddelli really are different species. We sequenced the 957 bp ND1 mitochondrial gene of 21 Saguinus monkeys (belonging to six species and nine morphotypes) and one Cebus monkey (the outgroup) and constructed phylogenetic trees using maximum parsimony, neighbor joining, and maximum likelihood methods. The phylogenetic trees obtained divided the genus Saguinus into two groups, one containing the small-bodied species S. fuscicollis and the other, the large-bodied species S. mystax, S. leucopus, S. oedipus, S. midas, S. bicolor. The most derived taxa, S. midas and S. bicolor, grouped together, while S. fuscicollis melanoleucus and S. f. weddelli showed divergence values that did not support the division of these morphotypes into subspecies. On the other hand, S. midas individuals showed divergence compatible with the existence of three subspecies, two of them with the same morphotype as the subspecies S. midas niger. The results of our study suggest that there is at least one Saguinus subspecies that has not yet been described and that the conservation status of Saguinus species and subspecies should be carefully revised using modern molecular approaches.
  • Association between reproductive traits and four microsatellites in Brangus-Ibagé cattle Animal Genetics

    Oliveira, João F.C. de; Neves, Jairo P.; Almeida, Eric A.; Steigleder, Clara Sabina; Moraes, José Carlos F.; Gonçalves, Paulo Bayard D.; Weimer, Tania A.

    Abstract in English:

    The aim of the present study was to verify associations between reproductive efficiency and four microsatellite markers located in synteny with genes involved in the regulation of reproductive mechanisms. A sample of 107 females from a Brangus Ibagé population (5/8 Aberdeen Angus x 3/8 Nelore) was characterized for ETH225 (D9S1) and MM12E6 (D9S20) microsatellites, mapped on chromosome 9, and HEL5 (D21S15) and AFZ1 (D21S37) on chromosome 21. Associations between the genetic markers and reproductive efficiency were determined by one-way analysis of variance using calving interval (CI), live weight at calving (LWC), live weight at first calving (LW1C) and live weight at second calving (LW2C) as dependent variables. The genotypes were classified according to allele size into homozygous for long alleles, homozygous for short alleles and heterozygous. A longer CI was observed for individuals homozygous for long alleles at the HEL5 locus compared with the others (p = 0.022). For the AFZ1 locus, an inverse correlation between allele size and calving interval was observed (p = 0.022), suggesting that homozygosity for long alleles at this microsatellite could be advantageous. Analysis of the combined effect of favorable genotypes at HEL5 and AFZ1 indicated that animals with unfavorable genotypes (homozygous for long alleles at HEL5 and homozygous for short alleles at AFZ1) presented a significantly longer CI (p = 0.003) when compared to the other genotypes. The ETH225 and MM12E6 systems did not present any association with CI. None of the systems studied showed any significant association with LWC, LW1C or LW2C.
  • Mitochondrial control region genetic diversity and maternal ancestry of a Brangus-Ibage cattle populations Animal Genetics

    Henkes, Luiz Ernani; Silva Jr, Wilson Araújo; Moraes, José Carlos Ferrugem; Weimer, Tania Azevedo

    Abstract in English:

    The genetic diversity of 277 nucleotides in the mitochondrial DNA control region (nt 15,964 to 16,240 in reference sequence) was analyzed in crossbreed beef cattle (Brangus-Ibage, 5/8 Bos primigenius taurus x 3/8 Bos primigenius indicus) as well as in some Nellore samples (B. p. indicus). Fifty-seven mutations were found in Brangus-Ibage comprising 18 haplotypes (haplotype diversity, h = 0.851 ± 0.041 and nucleotide diversity, ntd = 0.009 ± 0.006) and 66 in Nellore (h = 1.00 ± 0.27, ntd = 0.014 ± 0.012). These data indicated sequence identities of 99.6 and 92.1% between the B. p. taurus' reference sequence and Brangus-Ibage and Nellore, respectively. The comparison of our data with sequence data for 612 individuals recovered from GenBank showed a total of 205 haplotypes defined by 99 polymorphic sites. Most of the variability (53%) was due to differentiation within breeds. The phylogenetic tree constructed using the neighbor-joining method showed clearly the well-known dichotomy between B. p. taurus and B. p. indicus. The Brangus-Ibage clustered with B. p. taurus lineages; however, the displacement of Nellore from B. p. indicus branch probably indicates a substantial B. p. taurus maternal ancestry in some Nellore samples (obtained from GenBank) and reflects the primarily male-driven introduction of this breed in Brazil.
  • Dominance, epistasis, heritabilities and expected genetic gains Animal Genetics

    Viana, José Marcelo Soriano

    Abstract in English:

    Although epistasis is common in gene systems that determine quantitative traits, it is usually not possible to estimate the epistatic components of genotypic variance because experiments in breeding programs include only one type of progeny. As the study of this phenomenon is complex, there is a lack of theoretical knowledge on the contribution of the epistatic variances when predicting gains from selection and on the bias in estimating genetic parameters when fitting the additive-dominant model. The objective of this paper is to discuss these aspects. Regarding a non-inbred population, the genetic value due to dominance and the epistatic components of the genotypic value are not indicators of the number of favorable genes present in an individual. Thus, the efficiency of a selection process should be based on the narrow-sense heritability, a function only of additive variance. If there is no epistasis, generally it is satisfactory to assess the selection efficiency and to predict gain based on the broad-sense heritability. Regardless of the selection unit or type of epistasis, the bias in the estimate of the additive variance when assuming the additive-dominant model is considerable. This implies overestimation of the heritabilities at half sib family mean, plant within family and plant levels, and underestimation if the selection units are full sib progenies. The predicted gains will have a bias proportional to that of the heritability.
  • Estimation of genetic parameters for test-day milk yield in Holstein cows using a random regression model Animal Genetics

    Cobuci, Jaime Araujo; Euclydes, Ricardo Frederico; Lopes, Paulo Sávio; Costa, Claudio Napolis; Torres, Robledo de Almeida; Pereira, Carmen Silva

    Abstract in English:

    Test-day milk yield records of 11,023 first-parity Holstein cows were used to estimate genetic parameters for milk yield during different lactation periods. (Co)variance components were estimated using two random regression models, RRM1 and RRM2, and the restricted maximum likelihood method, compared by the likelihood ratio test. Additive genetic variances determined by RRM1 and additive genetic and permanent environmental variances estimated by RRM2 were described, using the Wilmink function. Residual variance was constant throughout lactation for the two models. The heritability estimates obtained by RRM1 (0.34 to 0.56) were higher than those obtained by RRM2 (0.15 to 0.31). Due to the high heritability estimates for milk yield throughout lactation and the negative genetic correlation between test-day yields during different lactation periods, the RRM1 model did not fit the data. Overall, genetic correlations between individual test days tended to decrease at the extremes of the lactation trajectory, showing values close to unity for adjacent test days. The inclusion of random regression coefficients to describe permanent environmental effects led to a more precise estimation of genetic and non-genetic effects that influence milk yield.
  • Analysis of restriction fragment length polymorphism in the kappa-casein gene related to weight expected progeny difference in Nellore cattle Animal Genetics

    Biase, Fernando H.; Garnero, Analia Del V.; Bezerra, Luiz A.F.; Rosa, Arthur J.M.; Lôbo, Raysildo B.; Martelli, Lúcia

    Abstract in English:

    Restriction fragment length polymorphism (RFLP) has been detected at the bovine kappa-casein locus. The polymorphism has been analyzed for its effects in cattle production, mostly for milk traits and even for maternal effect on pre-weaning weights. We used polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) to genotype 408 Nellore animals for the non-silent mutation (Thr/Ile136 and Asp/Ala148) that characterizes the A and B variants of the polymorphism and compared expected progeny difference (EPD) for a maternal effect on 120 and 210 days weights and direct EPD for 120, 210, 450 and 550 day weight between AA and AB animals. The EPD values were obtained from the University of São Paulo (Brazil) Nellore Cattle Breeding Program, which evaluated 266,272 animals in 2001. Analysis of Variance was used to compare weight expected progeny differences (EPDs) between animals genotyped as AA and AB. The A allele frequency was 0.911. Although the AA animals had higher weight EPDs than AB animals the differences were not statistically significant (p > 0.05).
  • Relationship between the Porcine Stress Syndrome gene and pork quality traits of F2 pigs resulting from divergent crosses Animal Genetics

    Band, Guilherme de Oliveira; Guimarães, Simone Eliza Facioni; Lopes, Paulo Sávio; Schierholt, Alex Sandro; Silva, Kleibe Moraes; Pires, Aldrin Vieira; Benevenuto Júnior, Augusto Aloísio; Gomide, Lúcio Alberto de Miranda

    Abstract in English:

    The PSS genotypes of 596 F2 pigs produced by initial mating of Brazilian commercial sows and native boars were characterized by PCR-RFLP and the pork quality traits were evaluated. Among the 596 pigs studied, 493 (82.7%) were NN and 103 (17.3%) were Nn. There were no differences between NN and Nn pigs in the following pork qualities: pHu (5.71 ± 0.16 vs 5.70 ± 0.11), intramuscular fat (1.55 ± 0.64% vs 1.65 ± 0.67%), shear force (5552 ± 878 g/1.2 cm vs 5507 ± 826 g/1.2 cm), lightness (44.96 ± 2.05 vs 45.01 ± 1.92), redness (0.64 ± 0.60 vs 0.79 ± 0.55), yellowness (6.62 ± 0.56 vs 6.65 ± 0.48), hue (84.28 ± 5.53 vs 83.41 ± 4.85), or chroma (6.68 ± 0.52 vs 6.73 ± 0.52). However, pork from Nn pigs had a significantly (p < 0.05) lower pH45 (6.41 ± 0.27 vs 6.51 ± 0.26) and greater drip (3.92 ± 1.90% vs 3.06 ± 1.60%), cooking (33.29 ± 2.26% vs 32.50 ± 2.54%) and total (35.67 ± 2.48% vs 34.01 ± 2.58%) loss compared to that of NN pigs. These results indicate that, even in divergent crosses, PSS gene carriers produce pork of poorer quality.
  • Relationship between the Porcine Stress Syndrome gene and carcass and performance traits in F2 pigs resulting from divergent crosses Animal Genetics

    Band, Guilherme de Oliveira; Guimarães, Simone Eliza Facioni; Lopes, Paulo Sávio; Peixoto, Jane de Oliveira; Faria, Danielle Assis; Pires, Aldrin Vieira; Figueiredo, Frederico de Castro; Nascimento, Carlos Souza do; Gomide, Lúcio Alberto de Miranda

    Abstract in English:

    The PSS genotypes of 596 F2 pigs produced by initial mating of Brazilian native boars commercial sows and were characterized by PCR-RFLP and their carcass and performance traits were evaluated. Among the 596 animals analyzed, 493 (82.72%) were characterized as NN and 103 (17.28%) as Nn. With respect to carcass traits, Nn animals presented higher (p < 0.05) right half carcass weight, left half carcass weight, loin depth and loin eye area, and lower shoulder backfat thickness, backfat thickness between last and next to last but one lumbar vertebrae and backfat thickness after last rib at 6.5 cm from the midline compared to NN animals. Nn animals also showed (p < 0.05) higher values for most of the cut yields, indicating higher cutting yields for animals carrying the n allele and lower values for bacon depth, confirming lower fat deposition in carcass. In addition, Nn animals presented (p < 0.05) lower values for the performance trait weight at 105 days of age. These results indicate that animals carrying the PSS gene generate leaner carcasses, higher cut yields, and that the effects of the gene can be observed even in divergent crosses.
  • Genetic trends in the performance and reproductive traits of pigs Animal Genetics

    Torres Filho, Rodolpho de Almeida; Torres, Robledo de Almeida; Lopes, Paulo Sávio; Pereira, Carmen Silva; Euclydes, Ricardo Frederico; Araújo, Cláudio Vieira de; Silva, Martinho de Almeida e

    Abstract in English:

    The selection and evaluation of breeds and purebred lines to be used as the parental stock is an important step for the success of any animal breeding program. In this study, the feed:gain ratio, backfat thickness corrected for 100 kg, age to 100 kg, average daily gain, and age of sow at first farrowing were used to evaluate the genetic trends for direct and maternal additive genetic values in Large White pigs. The total number of piglets born or born alive and the litter weight at birth were used as indicators of the genetic trends for the direct additive values. The genetic trends were calculated by regression of the average predicted genetic values per year for each trait versus the offspring's year of birth (for performance traits) or versus the dam year of birth (for reproduction traits). The genetic trend estimates for direct effects showed that selection decisions made during the breeding program effectively improved the performance traits. However, for reproductive traits, the regression estimates showed no definite trend. The genetic trends for the maternal effects were generally positive but of low magnitude.
  • Comparative cytogenetics of Hoplias malabaricus (Pisces, Erythrinidae): a population analysis in adjacent hydrographic basins Animal Genetics

    Vicari, Marcelo Ricardo; Artoni, Roberto Ferreira; Bertollo, Luiz Antonio Carlos

    Abstract in English:

    The chromosomes of specimens from four Hoplias malabaricus populations from headwaters of adjacent river basins at Ponta Grossa, southern Brazil, were investigated using differential staining techniques (C-banding, AgNO3 and CMA3) and fluorescent in situ hybridization (FISH) with an 18S rDNA probe. The diploid chromosome number in representatives of all four populations was invariably 2n = 42, with karyotypes composed of 12 pairs of metacentrics and 9 pairs of submetacentrics, without heteromorphic sex chromosomes. This kind of karyotype represents cytotype A in regard to cytotypes identified previously in H. malabaricus, exhibiting however, at the same time, some differences in the distribution of constitutive heterochromatin segments and in the locations of nucleolus organizer regions (NORs). The apparent karyotype similarity strongly suggests a close kinship among the studied populations, but the small differences detected in the examined chromosomal markers indicate some evolutionary divergence due to gene flow restriction among them.
  • A study of the constitutive heterochromatin and nucleolus organizer regions of Isocopris inhiata and Diabroctis mimas (Coleoptera: Scarabaeidae, Scarabaeinae) using C-banding, AgNO3 staining and FISH techniques Animal Genetics

    Bione, Edgar; Camparoto, Marjori Leiva; Simões, Zilá Luz Paulino

    Abstract in English:

    Meiotic and mitotic chromosomes of Isocopris inhiata and Diabroctis mimas were studied by standard staining procedures, C-banding, silver nitrate staining and FISH using Apis mellifera 28S rDNA as probe. Isocopris inhiata presented a 2n = 18 (8II+ Xy p) karyotype, composed of meta-submetacentric chromosomes with gradual reduction in size. The karyotype of D. mimas was 2n = 20 (9II+ Xy p), composed of meta-submetacentric (pairs 1, 2, 3, 4 and 7) and acrocentric (pairs 5, 6, 8 and 9) chromosomes, with gradual reduction in size. Analysis of constitutive heterochromatin revealed similar C-banding patterns for both species, showing pericentromeric and telomeric bands and diphasic chromosomes. In addition, the X chromosomes of these species were found to be almost completely heterochromatic. The presence of chromocenters was checked in one or more phases of prophase I of these species. All heterochromatin reacted positively for the silver stain. By FISH analysis we were able to locate the rDNA in medium-size autosome pairs in both species and in the X chromosome of D. mimas.
  • Use of the TSPY gene for sexing cattle Animal Genetics

    Lemos, Daniela Cristina; Rios, Álvaro Fabrício Lopes; Caetano, Lisandra Cristina; Lôbo, Raysildo Barbosa; Vila, Reginaldo Aparecido; Martelli, Lúcia; Takeuchi, Paula Lumy; Ramos, Ester Silveira

    Abstract in English:

    The Y-encoded, testis-specific protein (TSPY) is a Y-specific gene. The copy numbers of TSPY range from 20 to 60 in men and up to 200 in bulls. In this study, we examined the possibility of using the TSPY gene to sex cattle. DNA from blood samples of 100 Nelore cattle (50 males and 50 females) from the Nelore Cattle Breeding Program (PMGRN) was screened for TSPY by PCR using TSPY-specific primers. The assay was highly specific since all male samples were TSPY-positive and all female samples were negative. Positive results were also obtained at low DNA concentrations (less than 1 rhog/muL). These results showed that TSPY was a good male-specific marker, the usefulness of which was enhanced by the high copy number of the gene. This is the first report to demonstrate the applicability of TSPY for sexing cattle.
  • The use of PCR-RFLP as an identification tool for two closely related species of bats of genus Platyrrhinus Animal Genetics

    Ferreira, Juliana Machado; Martins, Felipe de Melo; Ditchfield, Albert; Morgante, João Stenghel

    Abstract in English:

    The bat species Platyrrhinus lineatus and P. recifinus (Phyllostomidae: Stenodermatinae) are ecologically important because of their capacity for seed dispersal. P. recifinus is endemic to the Atlantic rain forest and is considered vulnerable by the IUCN. The lack of distinct morphological features makes identification of the two species a difficult task. This study was aimed at testing the hypothesis that these are actually two distinct species by using PCR-RFLP of the mitochondrial cytocrome b gene. The results showed no shared haplotypes, demonstrating that these are, in fact, two distinct species. No polymorphism was obtained for P. recifinus, which could be a sign of low genetic diversity in this threatened species.
  • Cytogenetics and embryology of Eupatorium laevigatum (Compositae) Plant Genetics

    Bertasso-Borges, Maristela Sanches; Coleman, James Robert

    Abstract in English:

    Embryological studies indicate Eupatorium laevigatum to have Antennaria type diplospory with precocious embryony. The embryo sac is of the Polygonum type and the polar nuclei fuse before anthesis (maturation of the stamens). Endosperm development is autonomous and the central cell divides only after the initial stages of embryo formation. It is estimated that about 10% of the florets in anthesis contain an undivided egg which can be used for sexual reproduction. The study of microsporogenesis revealed abnormalities in chromosome pairing which result in the formation of univalents, bivalents, trivalents and higher polyvalents, with the consequent production of lagging chromosomes, unbalanced nuclei, micronuclei and sterile pollen. We found that, as represented by the material studied, E. laevigatum is an autohexaploid (2n = 6x = 60) in which each chromosome of a basic set of ten chromosomes is repeated six times and that E. laevigatum is an essentialy obligate apomictic.
  • Chromosomal organization and phylogenetic relationships in Hypochaeris species (Asteraceae) from Brazil Plant Genetics

    Ruas, Claudete de Fátima; Vanzela, André L.L.; Santos, Melissa O.; Fregonezi, Jeferson N.; Ruas, Paulo Maurício; Matzenbacher, Nelson I.; Aguiar-Perecin, Margarida L.R. de

    Abstract in English:

    The association of cytogenetic and molecular techniques has contributed to the analysis of chromosome organization and phylogeny in plants. The fluorochrome GC-specific CMA3, fluorescent in situ hybridization (FISH) and RAPD (Random Amplified Polymorphic DNA) markers were used to investigate chromosome structure and genetic relationships in Hypochaeris (Asteraceae). Seven species native to South America, and two species introduced from Europe (H. glabra and Hypochaeris sp) were studied. FISH with rDNA probes identified one or two loci of 18S-5.8S-25S rDNA in the South American Hypochaeris species and one locus in the European species. Only one 5S rDNA locus was seen in all species studied. Blocks of GC-rich heterochromatin (CMA-positive bands) associated to 18S-5.8S-25SrDNA loci were detected in all species investigated. Co-location of 5S rDNA and CMA bands was also observed, except for three South American species and Hypochaeris sp. In two South American species, additional CMA bands not related to rDNA were observed on the long arm of chromosome 2, near to the centromere. Hypochaeris glabra exhibited additional CMA-positive signals distributed at pericentromeric regions, on the short arms of all chromosomes. A total of 122 RAPD markers were used to determine the genetic relationships among species. The level of polymorphism was very high, revealing two genetic groups comprising the South American and the European species, thus supporting a previous hypothesis of monophyly of the South American Hypochaeris species. The coefficients of genetic similarity between European and South American species were 0.35, on average. Polymorphism was also high within the two groups. The genetic associations observed with RAPD markers were consistent with chromosome characteristics. Species carrying similar distribution of 45S rDNA loci and CMA-positive signals were included in the same group revealed by RAPDs. Cytogenetic and molecular data support the view that not only chromosome rearrangements, but also changes in DNA sequence took place during the diversification of the South American Hypochaeris species.
  • Identification of differentially expressed genes of Xanthomonas axonopodis pv. citri by representational difference analysis of cDNA Genetics Of Microorganisms

    Mehta, Angela; Rosato, Yoko B.

    Abstract in English:

    Xanthomonas axonopodis pv. citri is a phytopathogenic bacterium responsible for citrus canker, a serious disease which causes severe losses in citriculture around the world. In this study we report the differential expression of X. axonopodis pv. citri in response to specific treatments by using Representational Difference Analysis of cDNA (cDNA RDA). cDNAs from X. axonopodis pv. citri cultured in the presence of leaf extract of the host plant (Citrus sinensis), in vivo, as well as in the complex medium were hybridized against cDNA of the bacterium grown in the minimal medium. Sequencing of the difference products obtained after the second and third hybridizations revealed a total of 37 distinct genes identified by homology searches in the genome of X. axonopodis pv. citri. These genes were distributed in different functional categories, including genes that encode hypothetical proteins, genes involved in metabolism, cellular processes and pathogenicity, and mobile genetic elements. Most of these genes are likely related to growth and/or acquisition of nutrients in specific treatments whereas others might be important for the bacterium pathogenicity.
  • A shortcut in phage screening technique Genetics Of Microorganisms

    Andrade, Alexandre de; Teixeira, Paula Rezende; Siviero, Fábio; Santelli, Roberto Vicente

    Abstract in English:

    A simple modification of the traditional Benton & Davis technique for phage screening is presented that avoids the tedious sample dilutions of putative spots/phages towards the second screening. With the use of a sole agar plate and nylon filter, the modification distinguishes a true positive recombinant from a false positive, with high probability of success.
  • Evaluation of the genotoxic potential of the Hypericum brasiliense (Guttiferae) extract in mammalian cell system in vivo Mutagenesis

    Espósito, Anderson Victorino; Pereira, Danielli Maria Vieira; Rocha, Leandro Machado; Carvalho, José Carlos Tavares; Maistro, Edson Luis

    Abstract in English:

    Plants of the genus Hypericum, long used in folk medicine, contain active compounds which present, anti-septic, diuretic, digestive, expectorant, vermifugal, anti-depressive and other properties. The possible clastogenic effect of a H. brasiliense extract was tested in vivo on the bone marrow cells of Wistar rats. The extract was administered by gavage at doses of 50, 150 and 300 mg/kg body weight. Experimental and control animals were submitted to euthanasia 24 h after the treatment for micronucleus (MN) and chromosome preparations. H. brasiliense extract did not induce statistically significant increases in the average numbers of MN or chromosome aberrations in the test systems employed.
  • Chemoprotective effect of cysteamine against the induction of micronuclei by methyl methanesulfonate and cyclophosphamide Mutagenesis

    Santos-Mello, Renato; Deimling, Luiz Irineu; Lauer Júnior, Claudio; Carvalho, Thaís Rieger de

    Abstract in English:

    Cysteamine or 2-mercaptoethylamine (MEA) is an aminothiol with a well-known radioprotective action. No specific information is available in the literature about the possible chemoprotective action of MEA against genotoxic chemical agents. This paper presents the results of studies on the ability of MEA to protect mouse bone marrow polychromatic erythrocytes against the induction of micronuclei by alkylating agents such as methyl methanesulfonate (MMS) and cyclophosphamide (CP). We observed that MEA administered intraperitoneally 30 min before or 30 min after the administration of MMS or CP significantly reduced the frequency of micronucleated polychromatic erythrocytes (MNPCEs) induced by the alkylating agents. When MEA was administered in combination with MMS or CP the reduction in the frequency of MNPCEs did not reach statistically significant levels, although it reached values close to significance. With respect to the polychromatic erythrocyte/normochromatic erythrocyte (PCE/NCE) ratio, we observed that MEA did not provide significant protection against the bone marrow toxicity induced by CP.
  • Effects of H2O2, Fe2+ and Fe3+ on curcumin-induced chromosomal aberrations in CHO cells Mutagenesis

    Antunes, Lusânia Maria Greggi; Araújo, Maria Cristina Paiva; Dias, Francisca da Luz; Takahashi, Catarina Satie

    Abstract in English:

    The effects of H2O2, Fe2+ and Fe3+ on curcumin-induced clastogenicity were evaluated in CHO cells. Curcumin combined with H2O2 did not increase the chromosomal aberrations more than expected based on a simple additive effect. In contrast, the combination of curcumin-Fe significantly decreased the total number of chromosomal aberrations and the number of abnormal metaphases. The clastogenicity of curcumin may be related to its pro-oxidant properties and its ability to generate free radicals.
  • Population genetic structure of Brazilian shrimp species (Farfantepenaeus sp., F. brasiliensis, F. paulensis and Litopenaeus schmitti: Decapoda: Penaeidae) Evolutionary Genetics

    Gusmão, Jaqueline; Lazoski, Cristiano; Solé-Cava, Antonio M.

    Abstract in English:

    Penaeid shrimps are important resources for worldwide fisheries and aquaculture. In the Southwest Atlantic, Farfantepenaeus brasiliensis, F. paulensis, F. subtilis, Farfantepenaeus sp. and Litopenaeus schmitti are among the most important commercially exploited species. Despite their high commercial value, there is little information available on the different aspects of their biology or genetics and almost no data on their stock structure. We used allozymes to estimate variability levels and population genetic structure of F. brasiliensis, F. paulensis, L. schmitti and the recently detected species Farfantepenaeus sp. along as much as 4,000 km of Brazilian coastline. No population heterogeneity was detected in F. brasiliensis or L. schmitti along the studied area. In contrast, F ST values found for Farfantepenaeus sp. and F. paulensis indicate that the populations of those two species are genetically structured, comprising different fishery stocks. The largest genetic differences in F. paulensis were found between Lagoa dos Patos (South) and the two populations from Southeast Brazil. In Farfantepenaeus sp., significant differences were detected between the population from Recife and those from Fortaleza and Ilhéus.
  • Molecular markers in Chinese carps and their interspecific hybrids Evolutionary Genetics

    Fonteles, Soraia B.A.; Fernandes, Flora M. de C.; Lopes, Carlos E.; Kulikoski, Roberto; Kulikoski, Jussara M.; Almeida-Toledo, Lurdes F. de

    Abstract in English:

    A sample consisting of 2 Ctenopharyngodon idella (grass carp) females, 2 Aristichthys nobilis (bighead carp) males, and 10 interspecific hybrids was analyzed using a multidisciplinary approach. The 10 hybrids were the only surviving specimens of a crossbreeding experiment. The cytological approach, involving the measurement of the largest axis of the erythrocytes, revealed that the parental species were diploid, whereas, among the hybrids, eight individuals were triploid, and two were diploid. Two of the triploid individuals were analyzed cytogenetically and showed 2n = 3X = 72 chromosomes. Using the biochemical and molecular approaches, it was possible to confirm that one of the C. idella specimens was the true mother of the hybrids. Those data also allowed to rule out both males from parenthood. The molecular analyses also indicated the absence of gynogenetic individuals among the hybrids. We discuss the use of a multidisciplinary approach as an efficient tool in the monitoring of aquaculture programs.
  • Taxonomic implications of molecular studies on Northern Brazilian Teredinidae (Mollusca: Bivalvia) specimens Evolutionary Genetics

    Santos, Sonia Maria Lima; Tagliaro, Claudia Helena; Beasley, Colin Robert; Schneider, Horacio; Sampaio, Iracilda; Santos Filho, Carlos; Müller, Ana Cláudia de Paula

    Abstract in English:

    The current taxonomy of the Teredinidae (shipworms) is wholly based on morphology and up to now no molecular studies of the phylogeny of this group have been published. In the present study the relationships between four genera of the subfamilies Teredininae and Bankiinae were established and the efficiency of the 16S rRNA gene in characterizing four Teredinidae species was tested. Phylogenetic trees support the grouping of Bankia fimbriatula with Nausitora fusticula and of Neoteredo reynei with Psiloteredo healdi, but the genetic distances do not justify the classification of these species into two distinct subfamilies. The results show that B. fimbriatula, N. reynei and P. healdi specimens from the coast of the Brazilian state of Pará have five distinct 16S rRNA haplotypes, with one N. reynei haplotype differing from the other haplotypes in respect to at least seven sequences sites, indicating the existence of two very distinct sympatric lineages.
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